[Action mechanism of Mahuang Xixin Fuzi decoction for mice with influenza based on metabolomics information].

This study aimed to analyze the endogenous metabolite changes in the serum of mice infected with H1N1 virus after intervention by Mahuang-Xixin-Fuzi decoction (MXF) based on metabolomics method, investigate potential biomarkers and related metabolic pathways, and explore the therapeutic mechanism of MXF through metabolomics technology. Thirty-six Kunming (KM) mice were randomly divided into three groups: normal group, model group and MXF group. Influenza virus H1N1 was used by nasal drip to establish influenza mice model. The mice in MXF group were orally administrated with MXF for 6 consecutive days after inoculation, and the other two groups were given with equal volume of saline solution in the same way. Body weight, rectal temperature, morbidity and mortality were recorded daily. Serum samples were collected 24 hours after the last administration for HPLC-TOF-MS analysis. The results showed that as compared with the normal group, the body weight and rectal temperature were decreased in model group, and their lung index and mortality rate were significantly increased (P<0.05); MXF had good therapeutic effects on the abnormity of body weight, rectal temperature, lung index and high mortality rate of mice infected with H1N1 virus. The original data collected from the serum samples were analyzed with R language, MPP, SIMCA-P and other software, and significant changes were found in 14 kinds of endogenous substances from mice serum (P<0.05). As compared with model group, the potential metabolic markers in MXF group recovered to normal levels to a certain degree after being intervened by MXF. Further analysis with MetPA data platform showed that, the pathways involved in 14 metabolites included glucose metabolism, arachidonic acid metabolism, glycerophospholipids and sphingolipids metabolism etc. The metabolomics study and pharmacological experiment showed that MXF might play a role of efficacy by improving glucose metabolism, regulating arachidonic acid metabolism, glycerophospholipid and sphingolipid metabolic pathways.

[The Component Analysis of Petroleum Ether Extract in Scutellariae barbatae,Hedyotis diffusa and the Herb Pair and the Investigation of Anti-Endometrial Cancer Cells Activity].

Objective: To investigate the chemical components and the activity of anti-endometrial cancer cells of the petroleum ether extract in Scutellariae barbatae and Hedyotis diffusa and the herb pair. Methods: Main composition analysis and identification were determined by the GC-MS technology combined with Kovats retention index( KI). Activity of anti-endometrial cancer cells was researched by MTT assay. Results: Unsaturated fatty acid,esters,sterol and other compounds in Scutellariae barbatae,Hedyotis diffusa and the herb pair were identified by GC-MS. Hedyotis diffusa and the herb pair contained more anthraquinones which distinguished from Scutellariae barbatae. The IC50 values for HEC-1A cells of petroleum ether extract in Scutellariae barbatae and Hedyotis diffusa and the herb pair were 275. 204 µg / m L,105. 826 µg / m L,148. 645 µg / m L. The IC50 values for Ishikawa cells of petroleum ether extract in Scutellariae barbatae and Hedyotis diffusa and the herb pair are 189. 114 µg / m L,77. 974 µg / m L,137. 999 µg / m L. Conclusion: Petroleum ether extract in Scutellariac barbatae and Hedyotis diffusa and the herb pair has inhibition effect on the proliferation of HEC-1A and Ishikawa cells,the Hedyotis diffusa has the strongest activity of anti-endometrial cancer. It is speculated that the strongest activity could be related to the higher content of anthraquinones.

[GC-MS analysis for volatile components from alpiniae katsumadai semen by three extraction methods].

OBJECTIVE: To analysis the volatile components in Alpiniae Katsumadai Semen. METHODS: The volatile components were extracted from Alpiniae Katsumadai Semen by steam distillation, head space injection and supercritical fluid extraction respectively, and then analyzed by GC-MS combined with Kovat's retention index. RESULTS: The volatile components extracted by steam distillation or head space extraction were found more likely to be terpenoids, whereas components extracted by supercritical fluid extraction were more likely to be alkenes, alcohols and aromatic compounds. CONCLUSION: Different sample pre-treatment methods are focused on different types of volatile components; Identification of the volatile components by GC-MS combined with Kovat's retention index is more accurate and rapid.

Sterol regulatory element-binding protein-1/fatty acid synthase involvement in proliferation inhibition and apoptosis promotion induced by progesterone in endometrial cancer.

BACKGROUND: The number of endometrial cancer (EC) cases is escalating rapidly, with no evident improvements in survival rates. The downregulation of progesterone receptor, resulting in progestin resistance, is presently a major problem regarding the therapeutic aspect. On the basis of this, we can focus more on the downstream signaling pathways that are controlled by progesterone. Lipid biosynthesis mediated by sterol regulatory element-binding protein-1/fatty acid synthase (SREBP-1/FASN) is of utmost importance to the growth and the proliferation of EC cells, so we hypothesize that SREBP-1/FASN might be involved in suppressing the proliferation and promoting apoptosis in EC cells through the effects induced by progesterone. MATERIAL AND METHODS: The Cell Counting Kit-8 was used to analyze the growth inhibition ratio of Ishikawa cells upon treatment with megestrol acetate (MA; MA is a progesterone derivative, also known as 17α-acetoxy-6-dehydro-6-methylprogesterone) and to determine the 50% inhibitory concentration. Apoptosis ratio was analyzed by treatment of the cells with MA at 50% inhibitory concentration at different time intervals using Annexin V-FITC/propidium iodide. The protein and messenger RNA levels of SREBP-1 and FASN were compared between the experimental and control groups (MA-treated Ishikawa cells were considered to be the experimental group). RESULTS: The experimental group showed obvious growth inhibition that was time and concentration dependent. The apoptosis ratio was also significantly higher in the experimental group compared with the control group (P < 0.01). The protein and messenger RNA levels of SREBP-1 and FASN were significantly reduced by MA too. CONCLUSIONS: Sterol regulatory element-binding protein-1/FASN is involved in the proliferation suppression and apoptosis promotion brought about by MA in Ishikawa cells.

Three new C21 steroidal glycosides isolated from Metaplexis japonica and their potential inhibitory effects on tyrosine protein kinases.

Three new steroidal glycosides, metapregnoside A-C (II-IV), together with one known compound, byzantionoside B (I), were isolated from the fresh whole herb of Metaplexis japonica by using high-speed countercurrent chromatography and semi-preparative liquid chromatography. Their structures and relative configurations were elucidated by spectroscopic methods including 1D NMR, 2D NMR and HR-ESI-MS. The potential targets of compound I-IV were identified by virtual screening. And the potential inhibitory effects of these compounds on tyrosine protein kinases were compared by molecular docking. Byzantionoside B (I) was the first isolated compound from Metaplexis genus. The docking score of metapregnoside C (IV) was the highest. And the sugar chain residues at position C-20 in the pregn-4-en-3-one derivatives is the main factor affecting their docking scores on tyrosine protein kinases Fes/Fps.

A rapid and practical prediction method for the Arizona solvent system family used in high speed countercurrent chromatography.

Selecting the appropriate solvent system is the key to the successful separation of samples by using countercurrent chromatography. Although high-speed countercurrent chromatography has been widely used in the separation and preparation of natural products, the selection of a solvent system has always been a stumbling block to the application of high-speed countercurrent chromatography. In order to explore a rapid and practical prediction method to select countercurrent chromatography solvent system, five linear prediction models of the Arizona solvent system family (HEMW) was established by using fourteen compounds with different structures and five HPLC columns of different brands. And two different solvent system selection methods (The partition coefficient K of the target compound in the solvent system was in the range of 0.25 < K < 2.5) were proposed for targeted separation of compounds and multi-component separation in a complex sample respectively. The appropriate HSCCC solvent system of five known compounds was determined by a HPLC analysis and a shake flask test and the appropriate HSCCC solvent system of two Chinese herbal extracts was determined by a HPLC analysis to verify the prediction method. In this study, solid-liquid partition chromatography (HPLC) and liquid-liquid partition chromatography (HSCCC) were linked by polarity to simplify the screening process of solvent system. This method reduced the difficulty and workload of solvent system selection, which provided methods and ideas for more solvent system prediction models.

Single nucleotide polymorphism of SREBF-1 gene associated with an increased risk of endometrial cancer in Chinese women.

AIM: Elevated levels of sterol regulatory element-binding protein-1 (SREBP-1) have been found in endometrial cancer (EC), suggesting that it is essential to the development of EC. Obesity and diabetes have been established as known risk factors of EC, while SREBF-1 gene polymorphisms have also been found to be associated with obesity and type II diabetes. Therefore, we hypothesize that single nucleotide polymorphism (SNP) in SREBF-1 gene may be associated with increased risk of EC. METHOD: We analyzed the sequence of SREBF-1 in tissue samples from 30 EC cases and 6 benign controls using high throughput method. Based on the primary results, we selected one SNP (rs2297508) as a genetic marker to conduct a hospital-based case-control study with 139 EC cases and 129 benign controls. The samples were examined under the microscope to determine their histopathology prior to the SNP analysis using RT-PCR. RESULTS: Through sequence analysis, we found 10 SNPs of SREBF-1 associated with EC, including 3 new SNPs. Fourteen percent of EC showed the rs2297508 SNP with C allele, while only 7% had the C allele was present in benign controls (p = 0.027, OR = 1.983). Additionally, the C allele was associated with cancer differentiation (p<0.05) and the depth of myometrial invasion (p<0.05). CONCLUSION: Our study indicates that SNP (rs2297508) of SREBF-1 may serve as a genetic predisposition factor for the development of EC and screening of such genetic marker may be helpful in its early detection.